Biochemical characterization of a novel laccase from the basidiomycete fungus Cerrena sp. WR1

Sheng Chung Chen, Po Hung Wu, Yu Chang Su, Tuan Nan Wen, Yun Syuan Wei, Nai Chen Wang, Chih An Hsu, Andrew H.J. Wang, Lie Fen Shyur

研究成果: 雜誌貢獻文章同行評審

23 引文 斯高帕斯(Scopus)

摘要

This study reports a new white-rot fungus Cerrena sp. WR1, identified based on an 18S rDNA sequence, which can secrete extracellular forms of laccase with a maximal activity reaching 202 000 U l-1 in a 5-l fermenter. A laccase protein, designated Lcc3, was purified and shown to be N-linked glycosylated by PNGase F and liquid chromatography tandem mass spectrometry analyses. The respective full-length cDNA gene (lcc3) of the Lcc3 protein was obtained using polymerase chain reaction-based methods. Kinetic studies showed that the Km and kcat of the native Lcc3 were 3.27 μM and 934.6 s-1 for 2,2′-Azino-bis-(3-ethylbenzthiazoline-6- sulfonic acid), 849.1 μM and 147.9 s-1 for guaiacol, 392.7 μM and 109.2 s-1 for 2,6-dimethoxyphenol, and 881 μM and 115.5 s -1 for catechol, respectively. The Tm of Lcc3 was determined at 73.9°C and it showed a long t1/2 (120 min) at 50°C. The laccase was highly ethanol resistant, with 80 of its original activity was detected when incubated in 25 ethanol for 14 days. Furthermore, crude enzyme broth or Lcc3 could degrade lignin in kraft paper (26.5), and showed high decoloration efficiency (90) on synthetic dye Remazol Brilliant Blue R. Together, these data demonstrate that Cerrena sp. WR1 Lcc3 possesses novel biochemical and kinetic properties that may aid its application in industry.

原文英語
頁(從 - 到)761-769
頁數9
期刊Protein Engineering, Design and Selection
25
發行號11
DOIs
出版狀態已發佈 - 11月 2012
對外發佈

ASJC Scopus subject areas

  • 生物技術
  • 生物工程
  • 生物化學
  • 分子生物學

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